A Single Molecule Detection Platform (SMD) for a Leica SP8 TCS to analyse protein-protein interactions in living specimen.

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The discovery of the fluorescent protein GFP (green fluorescent protein) from certain jellyfish, has revolutionized our ability to study protein localization, protein dynamics and interactions of proteins. These and related fluorescent proteins allow us to investigate protein function within the complex environment of the living cell. Parallel to the developments in fluorescent protein biology, there have been advances in fluorescence imaging methods and microscopical systems that make it possible to localize proteins linked with such fluorescent proteins, to quantitate their abundance and to probe their mobility and interactions with each other. Recently, two imaging methods such as Förster resonance energy transfer (FRET) and fluorescence correlation spectroscopy (FCS) have been modified so that they can be done on user-friendly, commercially available laser scanning microscopes, replacing the need for complex custom-built microscopes. The combined advances in GFP biology and imaging methods are providing a massive stimulus for investigating the dynamic properties of proteins in living cells. The University of Exeter excels in many aspects of biomedical research, from fungal-related plant disease research to signalling biology in vertebrate embryonic development. In the past, biochemical approaches have been used to investigate protein-protein interactions. These observations can now be complemented and extended in real time in living cells. The latest generation of single molecule detectors is therefore a game changer for protein biology - in bacteria, fungi, plants and animals. It is now possible to generate dynamic maps of protein interactions in living cells using a fluorescence microscope. In this application, we seek support to purchase a modern single molecule detection platform (SMD), which will complement the existing facilities at the Exeter Bioimaging Centre.